Figure 2. Immunolocalization and Expression of cpLEPA.

A: Immunolocalization analysis of cpLEPA. The chloroplast, thylakoid, stroma and envelope fractions were subjected to immunoblot analysis with specific antisera against cpLEPA. Equal amounts of protein (20 µg) were loaded in each lane. The lanes marked cplepa-1, cplepa-2 and cplepa-1/35s::cpLEPA were loaded with equal amounts of total protein (20 µg). B: Salt washing of the membranes. The thylakoid membranes were incubated with 250 mM NaCl, 200 mM Na₂CO₃, 1 M CaCl₂ and 6 M urea for 30 min at 4°C. Then, the thylakoid proteins were separated by SDS-PAGE and immunoblotted with anti-LEPA, anti-RbcL (ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit) and anti-CP47 antibodies. RbcL and CP47 were used as markers. Thylakoid membrane preparations that had not been subjected to treatment were used as controls. C: Expression patterns of cpLEPA. Upper panel: cpLEPA expression levels in different organs of Arabidopsis, as determined by RT-PCR analysis. RNA samples isolated from seedlings, rosettes, flowers, roots, petiole, cauline tissue and siliques of wild-type plants were reverse-transcribed and subjected to PCR analysis. Middle panel: Transcript levels of cpLEPA in Arabidopsis leaves at 5, 15, 25, 35 and 45 d. Bottom panel: Light-induced accumulation of cpLEPA transcripts. Three-week-old plants grown under medium light (120 µmol m⁻² s⁻¹), low light (40 µmol m⁻² s⁻¹) or high light (500 µmol m⁻² s⁻¹) were used. ACTIN is shown as a control.